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결핵균 및 기타 3종 Mycobacteria 의 파쇄추출항원과 교차반응하는 폐결핵환자의 항체분석
황응수,조명제,심영수,김익상,차창용,국윤호,이승훈,한용철,배길한,김상재 대한미생물학회 1985 大韓微生物學會誌 Vol.20 No.1
It is important to discriminate between tuberculosis and tuberculosis-like disease by Mycobacteria other than tuberculosis in the serodiagnosis of tuberculosis. But because common antigens share among Mycobacteria, their antigenicities to human are similar. Therefore degree of cross-reactivity of anti-body in the sera of patients with tuberculosis between M. tuberculosis and Mycobacteria other than tu- berculosis should be checked to increase the specificity in the serodiagnosis of tuberculosis. The activity levels of IgG antibody in the sera of 106 patients confirmed as active pulmonary tuber-culosis and 30 normal healthy control person to the pressate extract antigen (TE, BE, AE, and FE antigen) from M. tuberculosis, M. bovis, M. avium, and M. fortuitum were measured by enzyme-linked immunosorbe:nt assay and the crossreactivity of IgG antibody with mycobacterial species was analysed. The results were as follows; 1. The activity level (O.D. at 492nm) of 1gG to TE antigen in sera of patients with pulmonary tuber- culosis was 0.228±0.167 in minimal tuberculosis; moderately advanced, 0.556±0.616; far advanced, 1.116±0.651 and 0.315±0.245 in miliary tuberculosis. 2. The actiivity level (O.D. at 492nm) of IgG to BE antigen in sera of patients with pulmonary tu-berculosis was 0.190+0.162 in minima1 tuberculosis; moderately advanced, 0.337+0.361; far advanced, 0.713+0.460 and 0.204±0.162 in miliary tuberculosis. 3. The activity level (O.D. at 492nm) of IgG to AE antigen in sera of patients with pulmonary tu-berculosis was 0.165±0.114 in minimal tuberculosis; moderately advanced, 0.392±0.494; far adven- ced, 0.751±0.512 and 0.2330.191 in miliary tuberculosis. 4. The activity level (O.D. at 492nm) of IgG to FE antigen in sera of patients with pulmonary tub- erculosis was 0.280±0.227 in minimal tuberculosis; moderately advanced, 0.460±0.564; far advan- ced, 0.845±0.573 and 0.257±0.103 in miliary tuberculosis. 5. The activity level (O.D. at 492nm) of IgG in sera of healthy control person was 0.126±0.084 to TE antigen, 0.105±0.041to BE antigen, 0.103±0.052 to AE antigen, and 0.095±0.061 to FE antigen. 6. Degree of correlation(r) in activity level of IgG between TE antigen and BE antigen was 0.905; between TE antigen and AE antigen, 0.760; between TE antigen and FE antigen, 0.790, and be- tween AE antigen and FE antigen,0.945. 7. As O.D. above 0.200 was determined positive for the serodiagnosis of pulmonary tuberculosis, the sensitivity and specificity in ELISA using TE antigen were 80% and 87% respectively, whereas in the case of using BE antigen, 66% and 100%; in the case of using AE antigen, 62% and 100 %, and in the case of using FE antigen, 72% and 93%, respecitively.
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황응수,김정헌,최은영,권예진,이동숙,황도영,정은숙 대한감염학회 2009 Infection and Chemotherapy Vol.41 No.1
Background : Porcine endogenous retroviruses (PERVs) form part of the chromosomes of all pigs. Since they can be produced as infectious virion and infect human cells, safety issues on PERVs infection to human are still controversial and is one of main hurdles of xenotransplantation using pig cells or organs. It has been reported that the established porcine cell line, PK-15, produces PERVs and can infect the human cell lines. Therefore, clonal analysis on human cell line infected with PERV is a prerequisite to characterize the infectivity to human cells and to investigate the harmfulness of PERVs to human. Materials and Methods : For the characterization of PERV that originates from porcine cell line, PK-15, full length PERV cloning from genomic DNA of PK-15 was performed and partial sequences of both ends were achieved. Cell clones from human cell line, 293, persistently infected with PERVs from PK-15 were established by the method of limiting dilution. Nested PCR and direct sequencing of PCR products in each clone were carried out so as to confirm the PERV genomes in each clone. The growth rate of each clone was checked using cell counting and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, the infectivity by reverse transcriptase (RT) assay, and genetic analysis by karyotyping. Results : A total of 12 genomic PERV clones could be retrieved; 1 with full length, 4 with defective forms, and others with irrelevant sequences. Intact PERV was thought to be able to infect 293 and the PERV-infected cell clones were selected by limiting dilution. PCR results confirmed that nine cell clones were infected with PERV, and sequence alignment data on PCR products of pol region from PK-15 and human cell clones with PERV showed very similar results. Cell counting and MTT assay for growth kinetics of each clone indicated that two clones showed reduced growth rate. However, it was difficult to verify the effect of PERV infection on the cell growth because of the presence of many genetic alterations in 293 parental cells. No RT activities were detected in the culture supernatant from PERV-infected 293 cell clones. Conclusion : The sequences of PERVs were detected in human cell clones after PERV infection, but PERV virions could not be detected from the culture supernatant by RT assay. Background : Porcine endogenous retroviruses (PERVs) form part of the chromosomes of all pigs. Since they can be produced as infectious virion and infect human cells, safety issues on PERVs infection to human are still controversial and is one of main hurdles of xenotransplantation using pig cells or organs. It has been reported that the established porcine cell line, PK-15, produces PERVs and can infect the human cell lines. Therefore, clonal analysis on human cell line infected with PERV is a prerequisite to characterize the infectivity to human cells and to investigate the harmfulness of PERVs to human. Materials and Methods : For the characterization of PERV that originates from porcine cell line, PK-15, full length PERV cloning from genomic DNA of PK-15 was performed and partial sequences of both ends were achieved. Cell clones from human cell line, 293, persistently infected with PERVs from PK-15 were established by the method of limiting dilution. Nested PCR and direct sequencing of PCR products in each clone were carried out so as to confirm the PERV genomes in each clone. The growth rate of each clone was checked using cell counting and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, the infectivity by reverse transcriptase (RT) assay, and genetic analysis by karyotyping. Results : A total of 12 genomic PERV clones could be retrieved; 1 with full length, 4 with defective forms, and others with irrelevant sequences. Intact PERV was thought to be able to infect 293 and the PERV-infected cell clones were selected by limiting dilution. PCR results confirmed that nine cell clones were infected with PERV, and sequence alignment data on PCR products of pol region from PK-15 and human cell clones with PERV showed very similar results. Cell counting and MTT assay for growth kinetics of each clone indicated that two clones showed reduced growth rate. However, it was difficult to verify the effect of PERV infection on the cell growth because of the presence of many genetic alterations in 293 parental cells. No RT activities were detected in the culture supernatant from PERV-infected 293 cell clones. Conclusion : The sequences of PERVs were detected in human cell clones after PERV infection, but PERV virions could not be detected from the culture supernatant by RT assay.
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황응수,박정규,차창용 대한면역학회 2004 Immune Network Vol.4 No.2
Viruses are obligate intracellular parasites which cause infection by invading and replicating within cells. The immune system has mechanisms which can attack the virus in extracellular and intracellular phase of life cycle, and which involve both non-specific and specific effectors. The survival of viruses depends on the survival of their hosts, and therefore the immune system and viruses have evolved together. Immune responses to viral infection may be variable depending on the site of infection, the mechanism of cell-to-cell spread of virus, physiology of the host, host genetic variation, and environmental condition. Viral infection of cells directly stimulates the production of interferons and they induce antiviral state in the surrounding cells. Complement system is also involved in the elimination of viruses and establishes the first line of defence with other non-specific immunity. During the course of viral infection, antibody is most effective at an early stage, especially before the virus enters its target cells. The virus- specific cytotoxic T lymphocytes are the principal effector cells in clearing established viral infections. But many viruses have resistant mechanism to host immune responses in every step of viral infection to cells. Some viruses have immune evasion mechanism and establish latency or persistency indefinitely. Furthermore antibodies to some viruses can enhance the disease by the second infection. Immune responses to viral infection are very different from those to bacterial infection. (Immune Network 2004;4(2):73-80) 바이러스는 세포 내 절대 기생체로서 세포에 침투하여 복제하여 증식한다. 면역계는 바이러스가 세포 밖에 존재하는 시기와 세포 내에 있는 시기 모두 공격을 할 수 있으며, 비특이적으로나 특이적인 반응을 보인다. 바이러스의 궁극적인 생존은 숙주의 생존 여부에 달려 있으므로 숙주의 면역체계와 바이러스는 상호 진화하여 왔다. 바이러스 감염에 대한 면역반응은 감염부위, 세포 간 바이러스의 전파기전, 숙주의 생리학적 상태, 유전적 소인과 환경요인에 따라 매우 다양하게 나타난다. 바이러스 감염은 인터페론의 생산을 유도해서 항바이러스 상태를 유발하고, 보체의 작용 등 선천면역에 의해 일차적으로 방어된다. 항체는 감염 초기 단계에 바이러스가 표적세포에 침투하기 전에는 매우 효과적으로 항바이러스 기능을 발휘한다. 확립된 바이러스 감염을 제거하고 완결시키는 데는 세포독성 T 림프구가 결정적인 역할을 한다. 그러나 바이러스는 이와 같은 단계별 숙주의 방어기전에 대항하는 기전을 갖고 있어서 바이러스에 따라서는 평생 숙주의 몸에서 잠복 또는 지속 감염을 이루게 된다. 한편 면역반응이 형성된 경우에 재감염이 되면 오히려 증상을 악화시키는 경우도 있는 등 바이러스 감염에 대한 면역반응의 특성은 다른 세균 등의 면역반응과 상당히 다른 점이 있다.
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황응수,조명제,차창룡,최강원,이승훈,장우현,Hwang, Eung-Soo,Cho, Myung-Je,Cha, Chang-Yong,Choe, Kang-Won,Lee, Seung-Hoon,Chang, Woo-Hyun The Korea Society for Microbiology 1986 大韓微生物學會誌 Vol.21 No.3
Serum samples from 51 patients with clinically suspected typhoid fever were tested for immunoglobulin G (IgG), IgM and IgA antibodies against the whole bacteria antigen of Salmonella typhi by an enzyme-linked immunosorbent assay. The levels of IgG and IgA antibody to-whole bacteria antigen were higher in the culture-proven patients than in controls. The levels of IgM antibody to- whole bacteria antigen showed better discrimination between culture negative patients and controls than those of IgG or IgA antibody to-whole bacteria antigen. The enzyme-linked immunosorbent assay was much more sensitive than the Widal test. It would be a useful tool for the diagnosis of typhoid fever with a single serum sample. 장티푸스가 의심되는 51명의 환자 혈청내에서 Salmonella typhi의 균체항원에 대한 IgG, IgM과 IgA 항체률 효소면역측정법으로 측정하였다. 균체항원에 대한 IgG와 IgA 항체가는 세균이 분리되어 장티푸스로 확진된 환자에서 건강대조군보다 높았다. 반면 세균을 분리하지는 못하였지만 임상증상 등으로 장티프스를 의심하는 환자에서는 IgM항체가는 건강대조군보다 높았다. 균체항원에 대한 항체를 효소면역 측정법으로 측정하는 것이 Widal 검사보다 민감도가 높았다. 장티푸스를 진단하는데 균체항원을 부착시켜 효소면역측정법을 사용하는 것이 유용할 것이다.
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